血管紧张素II对去甲肾上腺素释放上调金属蛋白酶2表达及主动脉夹层形成的影响
|
摘要
目的检验血管紧张素II通过促进去甲肾上腺素释放上调金属蛋白酶2(MMP-2)表达、参与主动脉夹层发病机制的假设。方法 36只SD大鼠随机分为对照组(12只,无预处理);洛沙坦预处理组[6只,洛沙坦40 mg/(kg·d)灌胃];交感神经化学消融组[6只,胍乙咤50μg/(g·d)皮下注射];多沙唑嗪组[6只,多沙唑嗪1 mg/(kg·d)灌胃];美托洛尔组[6只,美托洛尔100 mg/(kg·d)灌胃];两周后,获取大鼠主动脉环,用同位素标记去甲肾上腺素孵育大鼠主动脉环,记录各主动脉环基础去甲肾上腺素释放速度以及外源性血管紧张素II和/或去甲肾上腺素刺激后去甲肾上腺素释放速度,最后,ELISA法检测主动脉的MMP-2、MMP-9组织浓度。两肾一夹法建立血管紧张素升高模型,比较对照组[12只,生理盐水50μg/(g·d)皮下注射],两肾一夹组[18只,生理盐水50μg/(g·d)皮下注射],两肾一夹+交感消融组[18只,胍乙咤50μg/(g·d)皮下注射]。在第4、7和10周末处死动物并采用高效液相色谱法检测主动脉组织血管紧张素II,去甲肾上腺素水平,ELISA法检测主动脉组织MMP-2、MMP-9组织浓度。结果外源性血管紧张素II增加、去甲肾上腺素减慢交感神经末梢内源性去甲肾上腺素释放速度,MMP-2表达水平的变化与去甲肾上腺素释放的速度平行,而MMP-9不受到影响。交感神经消融预处理减弱血管紧张素II刺激的去甲肾上腺素释放增加、上调MMP-2的作用,而对MMP-9的表达没有影响;洛沙坦预处理降低了的去甲肾上腺素释放速度,MMP-2和MMP-9均有下调。多沙唑嗪和美托洛尔对各阶段去甲肾上腺素的释放速度均没有明显影响,但是美托洛尔在不影响去甲肾上腺素释放的情况下仍下调MMP-2的表达;多沙唑嗪和美托洛尔预处理对MMP-9表达没有明显影响。在体慢性实验表明,两肾一夹能升高主动脉的组织去甲肾上腺素水平和上调MMP-2,这种作用被交感神经化学消融减弱。结论主动脉交感神经末梢去甲肾上腺素释放速度调节主动脉金属蛋白酶2的表达,从而参与主动脉夹层的发病机制。
Objective To test the hypothesis that angiotensin II( Ang II) could enhance noradrenaline( NA) release from sympathetic nerve endings of the aorta thus contributing to the up-regulation of matrix metalloproteinase 2( MMP-2) during the formation of aortic dissection( AD). Methods Ang II,NA,MMP-2,MMP-9 of the aorta sample obtained during operation from aortic dissection patients were detected by High Performance Liquid Chromatography and ELISA and compared with controls. Isotope labelling method was used to test the impact of exogenous Ang II and noradrenaline on the NA release and MMP-2,MMP-9 expression on Sprague Dawley( SD) rat aorta rings in vitro. Two kidneys,one clip,models were replicated for further check of that impact in SD rats in vivo. Results The concentration of Ang II,MMP-2、9 was increased and NA concentration was decreased in aorta samples from AD patients. Exogenous Ang II enhanced while exogenous NA restrained NA release from aortic sympathetic endings. The Ang II stimulated NA release and the following MMP-2 up-regulation could be weakened by Losartan and chemical sympathectomy. Beta Mocker did not influence NA release but down-regulated MMP-2. Long term in vivo experiments confirmed that Ang II could enhance NA release and up-regulate MMP-2. Conclusion AD is initiated by MMP-2 overexpression as a result of increased NA release from sympathetic nervous endings in response to Ang II.
Objective To test the hypothesis that angiotensin II( Ang II) could enhance noradrenaline( NA) release from sympathetic nerve endings of the aorta thus contributing to the up-regulation of matrix metalloproteinase 2( MMP-2) during the formation of aortic dissection( AD). Methods Ang II,NA,MMP-2,MMP-9 of the aorta sample obtained during operation from aortic dissection patients were detected by High Performance Liquid Chromatography and ELISA and compared with controls. Isotope labelling method was used to test the impact of exogenous Ang II and noradrenaline on the NA release and MMP-2,MMP-9 expression on Sprague Dawley( SD) rat aorta rings in vitro. Two kidneys,one clip,models were replicated for further check of that impact in SD rats in vivo. Results The concentration of Ang II,MMP-2、9 was increased and NA concentration was decreased in aorta samples from AD patients. Exogenous Ang II enhanced while exogenous NA restrained NA release from aortic sympathetic endings. The Ang II stimulated NA release and the following MMP-2 up-regulation could be weakened by Losartan and chemical sympathectomy. Beta Mocker did not influence NA release but down-regulated MMP-2. Long term in vivo experiments confirmed that Ang II could enhance NA release and up-regulate MMP-2. Conclusion AD is initiated by MMP-2 overexpression as a result of increased NA release from sympathetic nervous endings in response to Ang II.
引文
[1]Wang XL,Ou L,Qin YW,et al.Association of the polymorphisms of MMP-9 and TIMP-3 genes with thoracic aortic dissection in Chinese Han population[J].Acta Pharmacol Sin,2014,35(3):351-355.
[2]Dab H,Hachani R,Hodroj W,et al.Interaction between sympathetic nervous system and renin angiotensin system on MMPs expression in juvenile rat aorta[J].Gen Physiol Biophys,2011,30(3):271-277.
[3]Salman IM,Sattar MA,Ameer OZ,et al.Role of norepinephrine&angiotensin II in the neural control of renal sodium&water handling in spontaneously hypertensive rats[J].Indian J Med Res,2010,131(6):786-792.
[4]王文靖,潘毅,杨涛等.两肾一夹型高血压大鼠模型的改良及评价[J].中国实验方剂学杂志,2012,18(1):203-205.
[5]Del Porto F,Cifani N,Proietta M,et al.MMP-12 and macrophage activation in acute aortic dissection[J].Cardiology,2014,128(4):314-315.
[6]Proietta M,Tritapepe L,Cifani N,et al.MMP-12 as a new marker of Stanford-A acute aortic dissection[J].Ann Mede,2014,46(1):44-48.
[7]Tsubaki M,Fujiwara D,Takeda T,et al.The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status,and statin-induced apoptosis is mediated via suppression of the Ras/ERK and Ras/m TOR pathways[J].Clin Exp Pharmacol Physiol,2017,44(2):222-234.
[8]Schulte K,Kumar M,Zajac JM,et al.Noradrenaline release in rodent tissues is inhibited by interleukin-1βbut is not affected by urotensin II,MCH,NPW and NPFF[J].Pharmacol Rep,2011,63(1):102-111.
[9]Gabriel SA,Antonangelo L,Capelozzi VL,et al.Hydrocortisone supresses inflammatory activity of metalloproteinase-8 in carotid plaque[J].Rev Bras Cir Cardiovasc,2015,30(3):295-303.
[10]Karpe PA,Gupta J,Marthong RF,et al.Insulin resistance induces a segmental difference in thoracic and abdominal aorta:differential expression of AT1 and AT2 receptors[J].J Hypertens,2012,30(1):132-146.
[11]Lu H,Rateri DL,Bruemmer D,et al.Involvement of the renin-angiotensin system in abdominal and thoracic aortic aneurysms[J].Clin Sci,2012,123(9):531-543.
[12]Xiong W,Meisinger T,Knispel R,et al.MMP-2 Regulates Erk1/2Phosphorylation and Aortic Dilatation in Marfan Syndrome[J].Circ Res,2012,110(12):e92-e101.
[13]Kurihara T,Shimizu-Hirota R,Shimoda M,et al.Neutrophil-derived matrix metalloproteinase 9 triggers acute aortic dissection[J].Circulation,2012,126(25):3070-3080.
[14]Dab H,Hachani R,Dhaouadi N,et al.Physiological regulation of MMPs and t PA/PAI in the arterial wall of rats by noradrenergic tone and angiotensin II[J].J Renin Angiotensin Aldosterone Syst.,2012,13(1):36-45.
[15]Rietz A,Spiers J.The relationship between the MMP system,adrenoceptors and phosphoprotein phosphatases[J].Br J Pharmacol,2012,166(4):1225-1243.
[2]Dab H,Hachani R,Hodroj W,et al.Interaction between sympathetic nervous system and renin angiotensin system on MMPs expression in juvenile rat aorta[J].Gen Physiol Biophys,2011,30(3):271-277.
[3]Salman IM,Sattar MA,Ameer OZ,et al.Role of norepinephrine&angiotensin II in the neural control of renal sodium&water handling in spontaneously hypertensive rats[J].Indian J Med Res,2010,131(6):786-792.
[4]王文靖,潘毅,杨涛等.两肾一夹型高血压大鼠模型的改良及评价[J].中国实验方剂学杂志,2012,18(1):203-205.
[5]Del Porto F,Cifani N,Proietta M,et al.MMP-12 and macrophage activation in acute aortic dissection[J].Cardiology,2014,128(4):314-315.
[6]Proietta M,Tritapepe L,Cifani N,et al.MMP-12 as a new marker of Stanford-A acute aortic dissection[J].Ann Mede,2014,46(1):44-48.
[7]Tsubaki M,Fujiwara D,Takeda T,et al.The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status,and statin-induced apoptosis is mediated via suppression of the Ras/ERK and Ras/m TOR pathways[J].Clin Exp Pharmacol Physiol,2017,44(2):222-234.
[8]Schulte K,Kumar M,Zajac JM,et al.Noradrenaline release in rodent tissues is inhibited by interleukin-1βbut is not affected by urotensin II,MCH,NPW and NPFF[J].Pharmacol Rep,2011,63(1):102-111.
[9]Gabriel SA,Antonangelo L,Capelozzi VL,et al.Hydrocortisone supresses inflammatory activity of metalloproteinase-8 in carotid plaque[J].Rev Bras Cir Cardiovasc,2015,30(3):295-303.
[10]Karpe PA,Gupta J,Marthong RF,et al.Insulin resistance induces a segmental difference in thoracic and abdominal aorta:differential expression of AT1 and AT2 receptors[J].J Hypertens,2012,30(1):132-146.
[11]Lu H,Rateri DL,Bruemmer D,et al.Involvement of the renin-angiotensin system in abdominal and thoracic aortic aneurysms[J].Clin Sci,2012,123(9):531-543.
[12]Xiong W,Meisinger T,Knispel R,et al.MMP-2 Regulates Erk1/2Phosphorylation and Aortic Dilatation in Marfan Syndrome[J].Circ Res,2012,110(12):e92-e101.
[13]Kurihara T,Shimizu-Hirota R,Shimoda M,et al.Neutrophil-derived matrix metalloproteinase 9 triggers acute aortic dissection[J].Circulation,2012,126(25):3070-3080.
[14]Dab H,Hachani R,Dhaouadi N,et al.Physiological regulation of MMPs and t PA/PAI in the arterial wall of rats by noradrenergic tone and angiotensin II[J].J Renin Angiotensin Aldosterone Syst.,2012,13(1):36-45.
[15]Rietz A,Spiers J.The relationship between the MMP system,adrenoceptors and phosphoprotein phosphatases[J].Br J Pharmacol,2012,166(4):1225-1243.